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目的:观察番茄红素(lycopene,LYC)对于血管内皮细胞功能的作用,探讨其作用机制。方法:人脐静脉内皮细胞(HUVECs)处理实验分组:对照组,H2O2组,H2O2+LYC组(1、2、4、8μmolL-1)。MTT法检测HUVECs存活率;免疫印迹法(Western blot)检测p38MAPK蛋白磷酸化水平、抗凋亡蛋白B淋巴细胞/白血病-2(bcl-2)及线粒体凋亡通路相关蛋白bax的表达;细胞黏附能力测定和伤口愈合实验检测HUVECs粘附率和迁移率;TUNEL法检测HUVECs凋亡率;ELASA法测定HUVECs内活性氧(ROS),超氧化物歧化酶(SOD),乳酸盐脱氢酶(LDH)释放量和caspase-3的活性。结果:H2O2损伤后HUVECs存活率显著降低(P<0.01),凋亡率显著增加(P<0.01),黏附和迁移能力显著降低(P<0.01),bax和p-p38MAPK的表达上调,bcl-2的表达下调,并且ROS、LDH的释放和caspase-3的活性增加(P<0.01),SOD的释放减少。而LYC的预处理可以明显逆转H2O2以上作用。结论:H2O2氧化应激损伤中,LYC保护内皮细胞可能与其抗过氧化损伤细胞凋亡,抑制异常的p38MAPK信号通路有关。
Objective: To observe the effect of lycopene (LYC) on the function of vascular endothelial cells and to explore its mechanism. Methods: Human umbilical vein endothelial cells (HUVECs) were divided into control group, H2O2 group, H2O2 + LYC group (1, 2, 4 and 8μmolL-1) The survival rate of HUVECs was detected by MTT assay. The phosphorylation of p38MAPK, the expression of bcl-2 and bax in mitochondrial apoptosis pathway were detected by Western blot. The adhesion and migration rates of HUVECs were tested by the ability test and wound healing test. The apoptosis rate of HUVECs was detected by TUNEL assay. The levels of reactive oxygen species (ROS), superoxide dismutase (SOD) and lactate dehydrogenase LDH release and caspase-3 activity. Results: The survival rate of HUVECs was significantly decreased (P <0.01), the apoptosis rate was significantly increased (P <0.01), the adhesion and migration ability was significantly decreased (P <0.01), the expression of bax and p- 2, and the release of ROS and LDH and the activity of caspase-3 increased (P <0.01), and the release of SOD decreased. However, pretreatment with LYC significantly reversed the effect of H2O2. Conclusion: In H2O2 oxidative stress injury, LYC protects endothelial cells may be related to its anti-peroxidative injury apoptosis, inhibition of abnormal p38MAPK signaling pathway.